Difference between revisions of "Team:UGent Belgium/LabNotebook"

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         <li class="list-group-item borderless"><b>August 29:</b> <p>We made a strong expression vector (pXS) by restriction of the linearized plasmid backbone pSB1C3 and insertion of gblock made up of promoter-RBS-cloning site-terminator-terminator (cut with EcoRI and PstI)</p>
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         <li class="list-group-item borderless"><b>August 29:</b> <p>
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         </li>           
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          <ul>
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            <li>
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We made a strong expression vector (pXS) by restriction of the linearized plasmid backbone pSB1C3 and insertion of gblock made up of promoter-RBS-cloning site-terminator-terminator (cut with EcoRI and PstI)
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            </li>
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            <li>
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              Our first attempts to isolate the inaZ gene from <i>P. syringae</i> and the inaX gene from <i>X. campestris</i>failed
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        <li class="list-group-item borderless"><b>August 30:</b> <p>
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          <ul>
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            <li>
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We transformed the strong expression vector (cf. 29/08) by electroshock into <i>E. coli</i> TOP10
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            </li>
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            <li>
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              As a backup strategy we also used CPEC with the backbone from the K584027 plasmid to generate a strong expression vector.
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          </ul>
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         </li>
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        <li class="list-group-item borderless"><b>August 31:</b> <p>
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          <ul>
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            <li>
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Second attempt to isolate the inaZ gene from <i>P. syringae</i> and the inaX gene from <i>X. campestris</i>. InaX failed, but inaZ was successful
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            </li>
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            <li>
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Transformation of the strong expression vector by restriction and ligation was not very successful, we only had a few colonies. The backbone amplification using CPEC was successful (cf 30/08).
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            </li>
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          </ul>
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        </li> 
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Revision as of 20:10, 6 October 2016

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Lab Notebook

August

  • August 29:

    • We made a strong expression vector (pXS) by restriction of the linearized plasmid backbone pSB1C3 and insertion of gblock made up of promoter-RBS-cloning site-terminator-terminator (cut with EcoRI and PstI)
    • Our first attempts to isolate the inaZ gene from P. syringae and the inaX gene from X. campestrisfailed
  • August 30:

    • We transformed the strong expression vector (cf. 29/08) by electroshock into E. coli TOP10
    • As a backup strategy we also used CPEC with the backbone from the K584027 plasmid to generate a strong expression vector.
  • August 31:

    • Second attempt to isolate the inaZ gene from P. syringae and the inaX gene from X. campestris. InaX failed, but inaZ was successful
    • Transformation of the strong expression vector by restriction and ligation was not very successful, we only had a few colonies. The backbone amplification using CPEC was successful (cf 30/08).